兔血浆异烟肼反相高效液相色谱分析及药代动力学研究

RP-HPLC Determination and Pharmacokinetics of Isoniazid in Rabbit Plasma

目的 建立兔血浆中异烟肼（Isoniazid,INH）的反相高效液相色谱分析方法（reversed-phase high-performance liquid chromatography,RP-HPLC）.研究其静脉给药后兔体内血浆药代动力学参数.方法 以Agilent ZORBAX SB-C18柱（5μm,4.6mm×250mm）为INH的分析柱,流动相为0.02 mol·L-1磷酸二氢钾缓冲液（H3PO4调pH4.0）：乙腈=55：45;流速1.0 mL·min-1;柱温：35C,血浆经桂皮醛衍生化后325nm波长下检测.8只新西兰大白兔分别按INH 11.43 mg·kg-1静滴,RP-HPLC法检测血浆药物浓度.所得血药浓度数据经DAS软件拟合分析.结果 INH在0.05～20μg·mL-1血浆浓度范围内线性关系良好（r=0.99989）,最低检测浓度为50ng·mL-1,平均提取回收率为100％,日内、日间变异系数（RSD =5）分别为1.2％ ～4.3％和2.5％ ～4.4％.INH在兔体内药代动力学过程符合双隔室模型（权重系数为1/cc）,T1/2=（1.92 ±0.80）h,Cmax=（9.70±1.38）mg·L-1,AUC（0-∞）=（20.97±3.11）mg· （h·L）-1.结论 本研究建立的兔血浆异烟肼HPLC方法具有较高的灵敏度、精密度和特异性,适用于兔体内药代动力学的研究.兔体内异烟肼药代动力学过程符合双隔室模型,体内药代谢动力学参数为异烟肼其它剂型的研究提供参考.

Objective To establish a assay method of reversed-phase high-performance liquid chromatography （RP-HPLC）,to determine plasmatic concentration of isoniazid （INH） and to explore the plasma Pharmacokinetics of INH in rabbit in vivo.Methods The chromatography conditions of INH were as followed：Agilent ZORBAX SB-C18 column （5 μm,4.6mm × 250mm）,0.02mol · L-1 phosphate buffer and acetionitrile （55：45,adjusted pH to 4.0 with H3PO4） as a mobile phase at 1.0 mL · min-1,column temperature at 35℃.The plasma was detected under 325 nm wavelength detection after derivatization.By cinnamic aldehyde 8 New Zealand white rabbits were injected with INH 11.43 mg · kg-1 intravenously,and the plasma drug concentration were analyzed by RP-HPLC assay.Plasma concentration data were fitted by DAS software.Results There was a good linear relationship within the range of 0.05 ～20μg · mL-1（r =0.99989）.The lowest concentration detected was 50 ng· mL-1.The extraction recovery was 100%.The precsion of intro-day and inter-day was 1.2% ～4.3% and 2.5% ～4.4%,respectively.INH pharmacokinetics in rabbits process was in line with double compartment model （weighting factor for the 1/cc）,T1/2 =（1.92 ± 0.80） h,Cmax =（9.70 ±1.38）mg · L-1,AUC（0-∞） =（20.97 ±3.11） mg · （h · L）-1 Conclusion This study established HPLC method with high sensitivity,precision and specificity for pharmacokinetic studies.INH pharmacokineties in rabbits process was in line with double compartment model and these findings can provide reference during pharmacokinetics investigations for the other preparations of INH.